We propose the Radial Scanning Objective (RSO), an innovative microscope objective that permits simultaneous automated neuron reconstruction using guided image collection, cell electrophysiological recording, and cell loading in live tissue. The RSO device will allow precise, accurate imaging by moving the objective lens relative to the tissue under study, without the need to move the electrodes, the tissue chamber, or the microscope itself. The RSO device and accompanying software technology will be the first commercially available product to a) enable direct recording of single distal synaptic connections formed between different areas, b) facilitate repeated monitoring of potential alterations in neuronal morphology during or as a result of stimulation, and c) permit simultaneous recording of one or more neurons formed between different areas in brain slices identified based on neuronal structure. This system will allow significant advancements in integrated studies of neural function and structure in basic neuroscience as well as in neurotoxicology and pharmacological research and development, representing clear progress beyond the state-of-the-art. PUBLIC HEALTH RELEVANCE: The proposed project will enable important new studies in neuroscience research that are not currently feasible. This project commercializes an innovative microscope hardware device and software combining the capabilities of guided automated neuron reconstruction, cell electrophysiological recording and cell filling in live tissue that will allow researchers to more accurately identify cells within a neural circuit, even those located at a distance from the original cell. The new device allows precise, accurate imaging by moving the objective lens relative to the tissue under study, without the need to move the electrodes, the tissue chamber, or the microscope itself. The overall effect of this project will open new horizons in integrated studies of neural function and structure in basic neuroscience as well as in neurotoxicology and pharmacological research and development and related fields involving research in neurological disease.